Design and Characterization of Rapid Optogenetic Circuits for Dynamic Control in Yeast Metabolic Engineering

Evan M. Zhao, Makoto A. Lalwani, Robert J. Lovelett, Sergio A. Garciá-Echauri, Shannon M. Hoffman, Christopher L. Gonzalez, Jared E. Toettcher, Ioannis G. Kevrekidis, José L. Avalos

Research output: Contribution to journalArticlepeer-review

Abstract

The use of optogenetics in metabolic engineering for light-controlled microbial chemical production raises the prospect of utilizing control and optimization techniques routinely deployed in traditional chemical manufacturing. However, such mechanisms require well-characterized, customizable tools that respond fast enough to be used as real-time inputs during fermentations. Here, we present OptoINVRT7, a new rapid optogenetic inverter circuit to control gene expression in Saccharomyces cerevisiae. The circuit induces gene expression in only 0.6 h after switching cells from light to darkness, which is at least 6 times faster than previous OptoINVRT optogenetic circuits used for chemical production. In addition, we introduce an engineered inducible GAL1 promoter (PGAL1-S), which is stronger than any constitutive or inducible promoter commonly used in yeast. Combining OptoINVRT7 with PGAL1-S achieves strong and light-tunable levels of gene expression with as much as 132.9 ± 22.6-fold induction in darkness. The high performance of this new optogenetic circuit in controlling metabolic enzymes boosts production of lactic acid and isobutanol by more than 50% and 15%, respectively. The strength and controllability of OptoINVRT7 and PGAL1-S open the door to applying process control tools to engineered metabolisms to improve robustness and yields in microbial fermentations for chemical production.

Original languageEnglish (US)
Pages (from-to)3254-3266
Number of pages13
JournalACS Synthetic Biology
Volume9
Issue number12
DOIs
StatePublished - Dec 18 2020

All Science Journal Classification (ASJC) codes

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

Fingerprint Dive into the research topics of 'Design and Characterization of Rapid Optogenetic Circuits for Dynamic Control in Yeast Metabolic Engineering'. Together they form a unique fingerprint.

Cite this